藤黃微球菌快速檢測(cè)方法的建立及rpf基因的表達(dá).pdf

1、湖南農(nóng)業(yè)大學(xué)碩士學(xué)位論文藤黃微球菌快速檢測(cè)方法的建立及rpf基因的表達(dá)姓名：陳宇明申請(qǐng)學(xué)位級(jí)別：碩士專(zhuān)業(yè)：基礎(chǔ)獸醫(yī)學(xué)指導(dǎo)教師：程天印20070601DevelopmentofrapiddetectiontechniqueandexpressionofrpfgeneofMicrococcusluteus’AbstractMierococcuslutens(formerlyMicrococcuslysodeikticus)isaspecia

2、lizedaerobicgramposifivebacteriaSkinisconsideredtobeaprimaryhabitatofthebacteriumandithasalsobeenwidelydetectedinwaterandsoilRecentlythisorganismwasrecognizedasanopportunisticpathogenandhasbeenimplicatedinnotonlyimmtmosu

3、ppressedpatientsbutalsoimmunosuppressedpigsandbirdsHowevertherese齟ehofpathogenicity、drugresistanceandassaysofdetectionislessInthispaperthefouraspectsWeredone：MicrococcusleteusisolatedfromtheYongzhouLiuyangLiLingChalingan

4、dotherfarmswereidentified；developmentofaPCRassaytodetectM1eteus；developmentofLAMPassaytodetectM1eteus；expressionofrpfgeneofM1eteusAroutinesystematicpathologicalcheckofporkerscausedbyaopportunisticpathogenWereconductedThe

5、mainpathologicalanatomicchangeswerechalactcrizedswellandhemorrhageintheliverintestinesandlungcouldalsobeenobservedThehistologicalpathologicalchangesWfflfedegenerationnecrosisinlivercellsandlymphnodeandinmuc：ousmembranece

6、llsofstomachandintestines；hyperemiaandhemorrhageinlung；PunetatehemorrhageinparenchumalorganssuchasliveLspleeninflammatorycellsinmosttissuewereusuallyobservedBasedonthe23SrRNAgenesequenceofMicrococcusluteusapdtrofspecific

7、primerswasdesignedtodetectM1utensThroughoptimizingtheamplifiedconditionafast、sensitiveandaccuratePCRassayWasdevelopedanditsaccuracyandsensitivitywereconfirmedThe1326bpspecificfragmentcouldbeamplifiedwithM1utensstandardSI

8、鞠JilbythisassaybutcouldnotbeamplifiedwithotherbacterialspeciessuchasEscherichiacoli，Salmonellatyphisuis、staphylococcusaldreRs，Streptococcus8ui$1058CFU／mLwasthelowestdetectinglimitformixedbacterialsolutionofM1uteusAhighly

9、specificsetoffourprimersF3、B3、FIP、BIPthatrecognizeatotalofsixdistinctsequences，Wasdesignedtotarget11251323bpregionofthe23SrRNAgenefromMieroeoecusluteusweoptimizedtheconditionsneededforthespecificamplificationofthe23SrRNA

10、geneandevaluatedthespecifictyansensitivityofthemethodTheseobservationsindicatethatnoamplificationofthetarget23SrRNAgenewasdetectedinotherrelatedbacteriawhentheLAMPprimerswereuseddetectionoftheclinicalsamplestheLAMPassayi